Fig. 2: GNF-2-deg and 2-12-2-deg induce CRBN- and proteasome-dependent depletion of DENV E in infected cells.

A Schematic of antiviral assay utilized in the study. B Western blots show concentration-dependent depletion of E in the presence of GNF-2-deg and 2-12-2-deg. Depletion of E is not observed for negative control compounds GNF-2-deg-BUMP or 2-12-2-deg-BUMP or when the experiment is conducted in CRBN-deficient cells. The representative results are shown from n = 3 independent experiments for GNF-2 derivative compounds in WT and n = 2 for all other conditions. C Schematic of experiments probing the mechanism of E degradation. Cells were infected with DENV2 at an MOI of 1 and treated with the E degrader (GNF-2-deg or 2-12-2-deg) along with the CRBN ligand lenalidomide or the neddylation inhibitor MLN4924 from 1 to 24 h post-infection. Due to cytotoxicity in the presence of proteasome inhibitor MG-132, cotreatment was limited to 21 to 24 h post-infection. Western blots show that depletion of E in the presence of GNF-2-deg and 2-12-2-deg is dependent on CRBN-binding (D), neddylation activity to activate CRL4^CRBN (E), and proteasome activity (F). Representative results of lenalidomide treatment are shown from n = 4 and n = 2 independent experiments for GNF-2 and 2-12-2 derivatives, respectively. Representative results of MLN4924 treatment are shown from n = 3 and n = 2 independent experiments for GNF-2 and 2-12-2 derivatives, respectively. Representative results of MG-132 treatment are shown from n = 2 independent experiments. Figures 2A and 2C were created with Biorender.com.